To track the anti-inflammatory substances in purslane,the lipopolysaccharide-induced RAW264.7 macrophage inflammation model was established,which was guided by the tracer of active substances.The extraction,separation and structural identification of anti-inflammatory substances in purslane were performed by column chromatography(for extraction),silica gel column chromatography(for separation),and preparative high performance liquid chromatography and gas chromatography-mass spectrometry(for analyses).The results showed that the three crude extracts obtained from purslane through sequential extractions with petroleum ether-ethanol,anhydrous ethanol and pure water solvents reduced the secretion of nitric oxide(NO)in the cells to 33.13,25.83 and 20.53 μmol/L,respectively,with the crude petroleum ether extract exhibiting the strongest inhibitory effect(P<0.05).The petroleum ether phase was further separated into four fractions,with the Fr.1,Fr.2 and Fr.3 fractions had stronger anti-inflammatory effects,though the Fr.1 and Fr.2 fractions contained potential toxic components.Therefore,the Fr.3 fraction was selected for further separation.The Fr.3 fraction was separated through a silica gel column to obtain three fractions.The Fr.3.1 subfraction exhibited the strongest inhibitory effect against the NO secretion(11.80 μmol/L).The Fr.3.1 subfraction was further purified by the preparative liquid chromatography and GC-MS analysis,and the main components of the Fr.3.1 subfraction were identified as stearic acid(47.09%),di(2-ethylhexyl)phthalate(13.21%)and other components.This study established a method for separating and purifying anti-inflammatory substances from purslane,and provides a theoretical reference for the development and utilization of purslane.
Portulaca oleracea L.anti-inflammatory activityextraction and isolationidentification
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